JP2021525625A - 神経調節技術 - Google Patents
神経調節技術 Download PDFInfo
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Abstract
Description
(A)絶食時:
(1)50mg/dL(2.8mmol/L)未満:インスリンショック;
(2)50mg/dLから70mg/dL(2.8mmol/Lから3.9mmol/L):低血糖または低血糖症;
(3)70mg/dLから110mg/dL(3.9mmol/Lから6.1mmol/L):正常;
(4)110mg/dLから125mg/dL(6.1mmol/Lから6.9mmol/L):上昇または障害(前糖尿病);および
(5)125mg/dL(7mmol/L):糖尿病。
(B)非絶食時(食後約2時間の食後):
(1)70mg/dLから140mg/dL(3.9mmol/Lから7.8mmol/L):正常;
(2)140mg/dLから199mg/dL(8mmol/Lから11mmol/L):上昇または境界線(前糖尿病);および
(3)200mg/dL(11mmol/L)以上:糖尿病。
神経調節が始まる前に、GE Vivid E9超音波システムおよび11Lプローブを超音波スキャンのために使用した。関心のある内部領域に対応する焦点領域を、動物の皮膚にラベル付けした。HIFUトランスデューサをラベル付けされた領域に配置した。HIFUトランスデューサの開口部に配置されたより小さな撮像プローブ(3S)を使用して、別の超音波スキャンも実行した。3Sプローブの撮像ビームを、HIFUビームと位置合わせした。したがって、HIFUビームが標的器官の画像(超音波スキャナで可視化)を使用して関心領域に向けられていることを確認することができた。
8から12週齢の成体雄のSprague−Dawleyラット(250〜300g;Charles River Laboratories)を、25℃で12時間の明暗サイクルにおいて飼育し、1週間順応させてから実験を行った。水と通常のげっ歯類の餌は自由に摂取できた。
エンドトキシン(Escherichia coliからのLPS、0111:B4;Sigma−Aldrich)を使用して、未処置の成体のSprague Dawleyラットに炎症および代謝機能障害(例えば、高血糖や高インスリン血症など)の重大な状態を引き起こした。LPSを、腹腔内(IP)注射によって動物(10mg/kg;Rosa−Ballinas PNAS、2008)に投与し、注射後4時間でピークに到達するが対照と比較して注射後8時間まで上昇したままであるTNFと循環グルコース濃度の大幅な上昇を引き起こした。印加された超音波エネルギーを使用した神経調節を、脾臓、右副腎、仙骨神経節、結節神経節、および/または孤束核で行った。LPS注射の前後1分間、超音波印加を行った。循環カテコールアミン濃度(例えば、ノルエピネフリンやドーパミン)の変化を分析するために、最後の超音波治療の15分後に血液サンプルを収集した。循環TNF濃度の変化を分析するために、最後の超音波治療の60分後に末端血液サンプルを収集した。血液サンプルは、サンプルの凝固を防ぐために抗凝固剤(二ナトリウム)EDTAとともに保存した。サンプルは、TNF(Lifespan)およびアセチルコリン(Lifespan)濃度の変化についてELISAアッセイによって分析した。カテコールアミン(例えば、ノルエピネフリンおよびドーパミン)濃度は、HPLC検出またはELISA(Rocky Mountain Diagnostic)分析を使用して評価した。
(A)動物を2〜4%イソフルランで麻酔した。
(B)動物は、処置中の高体温を防ぐために、水循環加温パッドに腹臥位で置いた。
(C)超音波刺激の標的関心領域の上方の領域(例えば、対象神経)を、刺激の前に使い捨てかみそりと動物用バリカンによって剃った。
(D)画像診断超音波を使用して、関心領域を空間的に選択した。
(E)その領域を、後で識別するために油性ペンでマークした。
(F)FUS超音波プローブまたはLogiQ E9プローブのいずれかを、画像診断超音波によって以前に識別された指定された関心領域に配置した。
(G)次に、超音波パルスを、単一の1分間のパルスを超えない単一の刺激の合計持続時間で実行した。超音波パルスのエネルギーは、熱損傷およびアブレーションまたはキャビテーションに関連するレベル(例えば、35W/cm2)には到達しない。
(H)そして、LPS(10mg/kg)を腹腔内注射することができる(急性または動態研究用)。あるいは、効果の持続期間のために、LPSは、代わりに後の指定された時点で注入してもよい。
(I)第2の1分間の超音波パルスを印加することができる。
(J)そして、動物は、急性研究(例えば、1時間)および動態研究(例えば、LPS後最大3時間まで変化する)のために、麻酔下でインキュベートすることができる。その後、動物を犠牲にし、組織と血液のサンプルを収集する。
切開は、腹膜腔の基部から始まり、胸膜腔まで行った。器官を迅速に取り出し、ホスファターゼ(0.2mMフェニルメチルスルホニルフルオリド、5μg/mLのアプロチニン、1mMベンズアミジン、1mMオルトバンデートナトリウムおよび2μMカンタリジン)とプロテアーゼ(Roche Diagnosticsに従い1μLから20mgの組織)阻害剤を含むPBS溶液でホモジナイズした。PBS溶液1mLあたり0.2g組織の標的最終濃度を全てのサンプルに適用した。血液サンプルは、サンプルの凝固を防ぐために抗凝固剤(二ナトリウム)EDTAとともに保存した。そして、サンプルは、分析まで−80℃で保存した。サンプルは、TNF(Lifespan/Abcam/ThermoFisher)濃度とアセチルコリン(Lifespan)濃度の変化についてELISAアッセイによって分析した。カテコールアミン濃度は、高速液体クロマトグラフィ(HPLC)検出または酵素結合免疫吸着測定法(ELISA)(Rocky Mountain Diagnostic)分析を使用して評価した。
血清サンプルは、前処理なしで直接HPLCに注入した。組織ホモジネートを最初に0.1M過塩素酸でホモジナイズし、15分間遠心分離した。次に、上澄みを分離し、サンプルをHPLCに注入した。
Claims (21)
- 対象の関心領域に機械的または超音波エネルギーを印加して、1つ以上の神経経路の調節を誘発することを含む方法であって、
前記関心領域が、結節神経節または仙骨神経節の少なくとも一部を含む、方法。 - 前記関心領域が結節神経節を含む、請求項1に記載の方法。
- 前記関心領域が仙骨神経節を含む、請求項1に記載の方法。
- 前記機械的または超音波エネルギーが超音波エネルギーである、請求項1に記載の方法。
- 前記関心領域が結節神経節を含み、前記1つ以上の神経経路が抗炎症経路およびグルコース調節経路である、請求項1に記載の方法。
- 前記関心領域が仙骨神経節を含み、前記1つ以上の神経経路が抗炎症経路およびドーパミン産生経路である、請求項1に記載の方法。
- 機械的または超音波エネルギーを関心領域に印加することが、前記対象内の遠位部位における1つ以上の目的分子の濃度の変化をもたらす、請求項1に記載の方法。
- 前記1つ以上の目的分子がグルコースを含む、請求項7に記載の方法。
- 前記1つ以上の目的分子がドーパミンを含む、請求項7に記載の方法。
- 前記1つ以上の目的分子が腫瘍壊死因子を含む、請求項7に記載の方法。
- 前記1つ以上の目的分子がアセチルコリンを含む、請求項7に記載の方法。
- 対象の関心領域に機械的または超音波エネルギーを印加して、2つ以上の神経経路の調節を誘発することを備える方法であって、
前記関心領域が、神経節の少なくとも一部を含む、方法。 - 前記2つ以上の神経経路が、神経免疫経路、抗炎症経路、ドーパミン産生経路、グルコース調節経路、インスリン産生経路、およびそれらの組み合わせから選択される、請求項12に記載の方法。
- 前記2つ以上の神経経路が、抗炎症経路およびグルコース調節経路を含む、請求項12に記載の方法。
- 前記2つ以上の神経経路が、抗炎症経路およびインスリン産生経路を含む、請求項12に記載の方法。
- 前記2つ以上の神経経路が、抗炎症経路およびドーパミン産生経路を含む、請求項12に記載の方法。
- システムであって、
1つ以上の神経経路を調節するために対象の関心領域に機械的または超音波エネルギーを印加するように構成されたエネルギー印加装置と、
コントローラであって、
前記関心領域を空間的に選択し、
前記エネルギー印加装置の1つ以上の調節パラメータを制御する
ように構成されたコントローラと
を備え、
前記関心領域が、結節または仙骨神経節の少なくとも一部を含む、システム。 - 前記エネルギー印加装置が、超音波トランスデューサである、請求項17に記載のシステム。
- 前記神経節が結節神経節を含む、請求項17に記載のシステム。
- 前記神経節が仙骨神経節を含む、請求項17に記載のシステム。
- 前記1つ以上の神経経路のうちの2つの神経経路が調節される、請求項17に記載のシステム。
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